Androgynomonoecious Jatropha curcas: Chromosomes, Isozymes, and Flowers Gender

Jatropha curcas (J. curcas) is usually monoecious plants, which have male and female flowers on the same inflorescence. However, J. curcas can be found as an androgynomonoecious plant (have male, female, and hermaphrodite flowers), even though very rare. Androgynomonoecious J. curcas can be identified after six months of planting when it had started flowering. Therefore, it is important to identify the characteristics of androgynomonoecious J. curcas that can differentiate between androgynomonoecious and monoecious plants in earlier stages of growth. The objectives of the research were to observe isozymes, chromosome and flowers gender of androgynomonoecious and monoecious J. curcas Banten and Lampung accessions. Seeds from five genotypes of J. curcas were used in the research. The observation was carried out on the chromosome and isozymes (Peroxidase and Esterase isozymes) could be used as markers to differentiate androgynomonoecious and monoecious plants. Observations about the flower gender from offsprings derived from different seeds were important to know the inheritance of flower gender. The androgynomonoecious and monoecious J. curcas were diploid with number of chromosomes 2n=2x=22. The chromosomes of androgynomonoecious have longer than that of monoecious J. curcas. The isozymes of androgynomonoecious J. curcas had four alleles and monoecious J. curcas (Banten female monoecious) had three alleles. The flower inflorescence and gender derived from androgynomonoecious plants were unstable, due to androgynomonoecious is intermediate state

Construction of RNA Interference Vector to Silence Aluminum Tolerance Gene Candidate in Rice cv Hawara Bunar

One of the aluminum (Al) tolerance gene candidates, namely B11 gene, has been successfully isolated from Al-tolerant rice cv Hawara Bunar. However, the role of the gene in Al tolerance in rice has not been known. RNA interference (RNAi) technique is an effective tool to examine the biological function of the target gene in plant. The objective of the research was to construct RNAi recombinant vector carrying untranslated region of the B11 gene. RNAi recombinant vector carrying 195 bp sized 3′UTR_B11 fragment as a double-stranded RNA (dsRNA) trigger has been successfully constructed using GATEWAY™ cloning technology, pENTR™/D-TOPO® as a shuttle vector, and pANDA vector as a destination vector. RNAi construct was successfully introduced into Agrobacterium tumefaciens AgL0, and has been infected to rice cv Hawara Bunar. Analysis of putative transgenic rice showed eight of 20 plants were transgenic carrying the B11-RNAi construct

Isolasi Dan Karakterisasi Gen Sitrat Sintase Bakteri Pseudomonas Aeruginosa Dari Filosfer Hevea Brasiliensis Muell. Arg.

Pseudomonas aeruginosa merupakan bakteri utama di dalam rizosfer yang mempunyai sifat-sifat yang dapat dimanfaatkan di dalam pertanian dan lingkungan. Bakteri tersebut mensekresikan asam organik yang dapat melepaskan fosfor dan melindungi akar dari keracunan aluminium. Sitrat merupakan asam organik yang dominan disekresikan oleh Pseudomonas di dalam tanah. Sitrat menujukkan afinitas terhadap aluminium dan menyediakan fosfor yang lebih tinggi dibandingkan asam organik lainnya. Asam organik ini disintesis dai sebuah reaksi antara aksaloasetat dan asetil KoA, dikatalisis oleh sitrat sintase (CS) di dalam siklus Kreb. Penelitian ini dilakukan untuk mengisolasi dan mengkarakterisasi sitrat sintase dari Pseudomonas aeruginosa yang telah diisolasi dari permukaan daun tanaman karet. Primer spesifik untuk gen CCS didesain berdasarkan sekuen gen sitrat sintase beberapa bakteri yang disimpan di Genbank. Primer tersebut digunakan untuk mengamplifikasi gen CS dengan menggunakan mesin PCR. Gen CS telah berhasil diisolasi dari bakteri filosfere Pseudomonas aerugunosa. Gen CS Pseudomonas aeruginosa (PaCS) tersebut terdiri dari 1287 pb dan menyandikan 428 asam amino. PaCS mempunyai kesamaan asam amino yang tinggi dan hidrofobisitas dengan CS bakteri lainnya dan diduga mempunyai persamaan aktivitas enzim. Diterima : 11 April 2013; Disetujui : 17 September 2013 How to Cite : Tistama, R., Widyastuti, U., & Suharsono. (2013). Isolasi dan karakterisasi gen sitrat sintase bakteri Pseudomonas aeruginosa dari filosfer Hevea Brasiliensis Muell. Arg.. Jurnal Penelitian Karet, 31(2), 127-138. Retrieved from http://ejournal.puslitkaret.co.id/index.php/jpk/article/view/14

ISOLATION AND CLONING OF cDNA OF GENE ENCODING FOR METALLOTHIONEIN TYPE 2 FROM MELASTOMA AFFINE

Metallothionein is an important protein for detoxifying heavy metal ions. This research was conducted to isolate and clone cDNA of gene encoding for metallothionein type 2 from Melastoma affine. Total RNA was isolated from young leaves. Total cDNA was synthesized from the total RNA by reverse transcription. The MaMt2 cDNA was successfully isolated by PCR technique. The MaMt2 cDNA was inserted into pGEM-T Easy and the recombinant plasmid was successfully introduced into Escherichia coli DH5α. DNA sequencing analysis showed that this cDNA is full length consisting of 246 pb encoding 81 amino acid residues. This cDNA is identical to mRNA of AtMt2 from Arabidopsis thaliana. It does not contain any restriction sites found in the cloning sites of pGEM-T easy. The deduced protein of MaMT2 contains 14 cysteine residues distributed in the Cys-Cys, Cys-X-Cys, and Cys-X-X-Cys motifs.   Key words: cDNA, metallothionein, Melastoma affine, cloning, cystein

Isolasi dan Respons Tumbuh Cendawan Mutualistik Akar pada Beberapa Tanaman Pangan dan Kehutanan

The study aims to isolate and test the effectiveness of mutualistic root symbiont fungi isolates from the roots of rubber plants grown in marginal acidic soil plantations in increasing the growth of food crops and forestry plants. The fungal were isolated by root surface sterilization methods. We obtained 19 fungal isolates consisting of 8 genera, namely Alternaria, Aspergillus, Cladosporium, Curvularia, Fusarium, Penicillium Paecilomyces, Trichoderma, and mycelia sterilia. All isolates were subjected to a pathogenicity test on the Centrosema pubescens plant. Five out of the 19 fungal isolates increased plant growth and showed no disease symptoms, and the Aspergillus section Nigri FKK 3 isolate showed the best response. The isolate was further analyzed to assess the growth response of food crops (rice and corn) and forestry plants (Acacia auriculiformis and Paraserianthes falcataria). The treatments consisted of 3 phosphate (P) concentrations, namely 20%, 50%, and 100% of the recommended field applications. The combination of mutualistic fungal inoculation of Aspergillus section Nigri FKK 3 and 50% P concentration exhibited the highest biomass growth response compared to other treatments. This finding can provide basic information for developing fungal-based fertilizers to increase the productivity of food crops and forestry plants on sub-optimal land.   Keywords: food crops, phosphate fertilizer, forestry trees, plant growth improvement, root mutualistic fung

ISOLASI DAN KARAKTERISASI BAKTERI DENITRIFIKASI SEBAGAI AGEN BIOREMEDIASI NITROGEN ANORGANIK

Denitrifikasi merupakan salah satu proses utama yang mengurangi kandungan senyawa nitrogen anorganik di lingkungan. Proses ini dapat digunakan untuk mengatasi kelebihan senyawa nitrogen anorganik yang tinggi di kolam budidaya perikanan. Penelitian ini bertujuan untuk mengisolasi dan mengkarakterisasi isolat bakteri denitrifikasi sebagai agen bioremediasi senyawa nitrogen anorganik. Sebanyak 21 isolat bakteri pereduksi nitrat berhasil diisolasi dari medium pengkayaan dengan konsentrasi nitrat 100 µM dan 1500 µM. Sebanyak 6 isolat merupakan kelompok bakteri denitrifikasi (fermentatif negatif) dan 15 isolat termasuk kelompok bakteri fermentatif. Berdasarkan hasil seleksi didapatkan isolat HNF5 dan LNF mempunyai kemampuan reduksi nitrat yang tinggi. Aktivitas reduksi nitrat terjadi dari awal inkubasi, di mana aktivitas paling cepat terjadi pada fase eksponensial pertumbuhan bakteri. Isolat HNF5 dan LNF memiliki kecepatan maksimum reduksi nitrat (Vmaks) 0,17 mM.h-1 dan 0,16 mM.h-1 dengan nilai konstanta Michaelis-Menten (Km) 0,40 mM dan 0,28 mM. Identifikasi dengan sekuen 16S-rRNA memperlihatkan bahwa isolat HNF5 dan LNF mempunyai kemiripan dengan Pseudomonas aeruginosa

Introduksi Gen Sitrat Sintase Pseudomonas aeruginosa ke Nicotiana tabacum dan Jatropha curcas untuk Meningkatkan Toleransi terhadap Cekaman Aluminium

Cekaman aluminium merupakan faktor penghalang utama untuk pengembangan komoditas tanaman unggulan di lahan-lahan masam. Sintesis dan sekresi asam organik oleh akar merupakan salah satu mekanisme mengurangi cekaman aluminium. Penelitian ini bertujuan mengintroduksikan gen sitrat sintase bakteri P. aeruginosa (PaCS) ke dalam tanaman tembakau (N. tabacum) dan Jatropha curcas. PaCS berhasil diintroduksi dengan batuan Agrobacterium tumefaciens baik ke dalam jaringan tembakau maupun J. curcas.. Toleransi tembakau transgenik ditandai dengan pertambahan panjang akar yang lebih tinggi dibanding tipe liarnya pada cekaman 0,3 mM AlCl3. Toleransi tembakau transgenik dapat mengurangi masuknya Al ke dalam jaringan akar. Sifat toleran gen PaCS diturunkan mengikuti Hukum Mendel. Persentase introduksi gen PaCS ke dalam J. curcas hanya 1% dari jumlah eksplan yang ditranformasi. Kata kunci : introduksi gen; sitrat sintase; cekaman aluminium; N. tabacum; J. curca

Keragaman Genetik Kelapa Sawit (Elaeis Guineensis Jacq.) Asal Angola Menggunakan Marka SSR

Effort to increase productivity and other elite characters in Indonesia oil palm breeding program is facing a problem because of the narrow genetic diversity. To broaden the genetic diversity, germplasm exploration has been done in Angola, Central Africa. The objective of this research was to assess the genetic diversity and population structure of Angola originated oil palm germplasm based on 20 SSR markers. The plant materials used were 27 accessions consisted of 136 palms planted in Riau, Sumatera. The DNA was isolated and amplified using PCR. Phylogeny analysis was constructed using Unrooted Neighbor-Joining by DARwin software 6.0.8. The result showed that polymorphic information content (PIC) value is 0.55 (0.17 to 0.75 for each locus) with 102 total number of alleles. Genetic diversity between individuals was higher compared to the genetic diversity within accessions or regions and between accessions or regions. Phylogenetic analysis of 27 accessions showed that accessions were divided into three main groups. Every group containing individuals originated from 5 spatial distribution regions. Principal coordinates analysis (PCoA) showed that accessions were distributed in one structure. Using more primers and samples to get more representative data is recommended for the following research

RNAi dari Fragmen 3’UTR Gen Penyandi H+ -ATPase Membran Plasma Melastoma malabathricum L. dapat Menghambat Pertumbuhan Tanaman Tersebut

The RNA silencing technique is an effective tool to examine the biological function of the target mRNA in plants. The recent development of GATEWAYTM cloning technology makes it easy to construct the RNAi vectors with trigger sequences and to analyze the function of a target gene. The objective of this research was to construct RNAi including the 3’UTR fragment of the gene coding plasma membrane H+-ATPase from Melastoma malabathricumL., 3’UTRMmpma. RNAi vector had been successfully constructed using GATEWAYTM cloning technology with the 3’UTRMmpma was used as double-stranded RNA (dsRNA) trigger sequence, pENTRTM/D-TOPO®as entry vector, and pANDA plasmid as destination vector. RNAi had been successfully introduced into M. malabathricumL. mediated by A. tumefaciensEHA101 to analyze the function of Mmpma gene in the detoxifying Al stress. Based on the test of transgenic plants tolerance to Al stress showed that in the nutrient solution including 3.2 mM Al (AlCl3.6H2O), the transgenic plants underwent growth suppression especially roots and leaves, whereas non-transgenic plants underwent growth normally. It showed that suppression of Mmpmagene expression by RNAi to M. malabathricumL. caused the plant became sensitive to Al.Keywords: 3’UTRMmpma, A. tumefaciens, Al stress, RNAi vecto